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An increase in confirmed human salmonellosis cases in the EU after 2014 triggered investigation of contributory factors and control options in poultry production. Reconsideration of the five current target serovars for breeding hens showed that there is justification for retaining Salmonella  Enteritidis, S almonella  Typhimurium (including monophasic variants) and Salmonella  Infantis, while Salmonella Virchow and Salmonella  Hadar could be replaced by Salmonella  Kentucky and either Salmonella Heidelberg, Salmonella  Thompson or a variable serovar in national prevalence targets. However, a target that incorporates all serovars is expected to be more effective as the most relevant serovars in breeding flocks vary between Member State (MS) and over time. Achievement of a 1% target for the current target serovars in laying hen flocks is estimated to be reduced by 254,400 CrI 95 [98,540; 602,700] compared to the situation in 2016. This translates to a reduction of 53.4% CrI 95 [39.1; 65.7] considering the layer-associated human salmonellosis true cases and 6.2% considering the overall human salmonellosis true cases in the 23 MSs included in attribution modelling. A review of risk factors for Salmonella in laying hens revealed that overall evidence points to a lower occurrence in non-cage compared to cage systems. A conclusion on the effect of outdoor access or impact of the shift from conventional to enriched cages could not be reached. A similar review for broiler chickens concluded that the evidence that outdoor access affects the occurrence of Salmonella is inconclusive. There is conclusive evidence that an increased stocking density, larger farms and stress result in increased occurrence, persistence and spread of Salmonella in laying hen flocks. Based on scientific evidence, an impact of Salmonella control programmes, apart from general hygiene procedures, on the prevalence of Campylobacter in broiler flocks at the holding and on broiler meat at the end of the slaughter process is not expected., (© 2019 European Food Safety Authority. EFSA Journal published by John Wiley and Sons Ltd on behalf of European Food Safety Authority.)

The qualified presumption of safety (QPS) procedure was developed to provide a harmonised generic pre-evaluation to support safety risk assessments of biological agents performed by EFSA's Scientific Panels. The taxonomic identity, body of knowledge, safety concerns and antimicrobial resistance were assessed. Safety concerns identified for a taxonomic unit are, where possible and reasonable in number, reflected by 'qualifications' which should be assessed at the strain level by the EFSA's Scientific Panels. During the current assessment, no new information was found that would change the previously recommended QPS taxonomic units and their qualifications. Between April and September 2018, the QPS notification list was updated with 48 microorganisms from applications for market authorisation. Of these, 30 biological agents already had QPS status, 15 were excluded from the QPS exercise by the previous QPS mandate (five filamentous fungi) or from further evaluations within the current mandate (two notifications of Enterococcus faecium , one of Streptomyces spp. and seven of Escherichia coli ). One taxonomic unit was (re)evaluated: Pseudomonas fluorescens had been previously evaluated in 2016, and was now re-evaluated within this mandate. The revision of the literature supports the previously identified safety concerns (e.g. production of biocompounds with antimicrobial activity and virulence features), preventing the inclusion of P. fluorescens in the QPS list. Mycobacterium setense and Komagataeibacter sucrofermentans were evaluated for the first time. M. setense cannot be considered for the QPS assessment because there are significant safety concerns. K. sucrofermentans ( Acetobacter xylinus subsp. sucrofermentans ) can be proposed for the QPS list but only for production purposes. The QPS status of Corynebacterium glutamicum is confirmed with the qualification extended to other production purposes., (© 2019 European Food Safety Authority. EFSA Journal published by John Wiley and Sons Ltd on behalf of European Food Safety Authority.)

Parasites are important food-borne pathogens. Their complex lifecycles, varied transmission routes, and prolonged periods between infection and symptoms mean that the public health burden and relative importance of different transmission routes are often difficult to assess. Furthermore, there are challenges in detection and diagnostics, and variations in reporting. A Europe-focused ranking exercise, using multicriteria decision analysis, identified potentially food-borne parasites of importance, and that are currently not routinely controlled in food. These are Cryptosporidium spp., Toxoplasma gondii and Echinococcus spp. Infection with these parasites in humans and animals, or their occurrence in food, is not notifiable in all Member States. This Opinion reviews current methods for detection, identification and tracing of these parasites in relevant foods, reviews literature on food-borne pathways, examines information on their occurrence and persistence in foods, and investigates possible control measures along the food chain. The differences between these three parasites are substantial, but for all there is a paucity of well-established, standardised, validated methods that can be applied across the range of relevant foods. Furthermore, the prolonged period between infection and clinical symptoms (from several days for Cryptosporidium to years for Echinococcus spp.) means that source attribution studies are very difficult. Nevertheless, our knowledge of the domestic animal lifecycle (involving dogs and livestock) for Echinoccocus granulosus means that this parasite is controllable. For Echinococcus multilocularis , for which the lifecycle involves wildlife (foxes and rodents), control would be expensive and complicated, but could be achieved in targeted areas with sufficient commitment and resources. Quantitative risk assessments have been described for Toxoplasma in meat. However, for T. gondii  and  Cryptosporidium  as faecal contaminants, development of validated detection methods, including survival/infectivity assays and consensus molecular typing protocols, are required for the development of quantitative risk assessments and efficient control measures., (© 2018 European Food Safety Authority. EFSA Journal published by John Wiley and Sons Ltd on behalf of European Food Safety Authority.)

In 2017, EFSA published a 'simplified' food safety management system (FSMS) for certain small retail establishments (butcher, grocer, baker, fish and ice cream shop) based on the application of prerequisite programme (PRP) criteria. The aim of this opinion was to develop similar FSMSs for other small retail enterprises including retail distribution centres, supermarkets, restaurants (including pubs and other catering activities) and food donation. The latter presents several novel food safety challenges because donated food may be nearing the end of its shelf-life and several actors are involved in the food donation chain, each reliant on each other to assure food safety. In this opinion, the simplified approach to food safety management is presented based on a fundamental understanding of processing stages and the activities contributing to increased occurrence of the hazards (biological, chemical (including allergens) or physical) that may occur. Control is achieved using PRP activities as previously described but with a modified 'temperature control' PRP and the addition of PRPs covering shelf-life control, handling returned foods, shelf-life evaluation for food donation, allocation of remaining shelf-life, and freezing food intended for donation. Examples of the simplified approach are presented for retail distribution centres, supermarkets, restaurants and food donation., (© 2018 European Food Safety Authority. EFSA Journal published by John Wiley and Sons Ltd on behalf of European Food Safety Authority.)

In the past, the horizontal transfer of antimicrobial resistance genes was mainly associated with conjugative plasmids or transposons, whereas transduction by bacteriophages was thought to be a rare event. In order to analyze the likelihood of transduction of antimicrobial resistance in the field of clinical veterinary medicine, we isolated phages from Escherichia coli from a surgery suite of an equine clinic. In a pilot study, the surgery suite of a horse clinic was sampled directly after surgery and subsequently sampled after cleaning and disinfection following a sampling plan based on hygiene, surgery, and anesthesia. In total, 31 surface sampling sites were defined and sampled. At 24 of these 31 surface sampling sites, coliphages were isolated. At 12 sites, coliphages were found after cleaning and disinfection. Randomly selected phages were tested for their ability of antimicrobial resistance transduction. Ten of 31 phages were detected to transfer antimicrobial resistance. These phages most often transduced resistance to streptomycin, encoded by the addA1 gene ( n = 9), followed by resistance to chloramphenicol by cmlA ( n = 3) and ampicillin ( n = 1). This is, to the best of our knowledge, the first report on antimicrobial resistance-transferring bacteriophages that have been isolated at equine veterinary clinics.

Echo Planar Imaging (EPI) is most commonly applied to acquire diffusion-weighted MR-images. EPI is able to capture an entire image in very short time, but is prone to distortions and artifacts. In diffusion-weighted EPI of the kidney severe distortions may occur due to intestinal gas. Turbo Spin Echo (TSE) is robust against distortions and artifacts, but needs more time to acquire an entire image compared to EPI. Therefore, TSE is more sensitive to motion during the readout. In this study we compare diffusion-weighted TSE and EPI of the human kidney with regard to intravoxel incoherent motion (IVIM) and diffusion tensor imaging (DTI). Images were acquired with b-values between 0 and 750s/mm 2 with TSE and EPI. Distortions were observed with the EPI readout in all volunteers, while the TSE images were virtually distortion-free. Fractional anisotropy of the diffusion tensor was significantly lower for TSE than for EPI. All other parameters of DTI and IVIM were comparable for TSE and EPI. Especially the main diffusion directions yielded by TSE and EPI were similar. The results demonstrate that TSE is a worthwhile distortion-free alternative to EPI for diffusion-weighted imaging of the kidney at 3Tesla., (Copyright © 2017. Published by Elsevier GmbH.)

Background: MRI of synovitis relies on use of a gadolinium-based contrast agent. Diffusion-weighted MRI (DWI) visualises thickened synovium but is of limited use in the presence of joint effusion., Objective: To investigate the feasibility and diagnostic accuracy of diffusion-weighted MRI with intravoxel incoherent motion (IVIM) for diagnosing synovitis in the knee joint of children with juvenile idiopathic arthritis., Materials and Methods: Twelve consecutive children with confirmed or suspected juvenile idiopathic arthritis (10 girls, median age 11 years) underwent MRI with contrast-enhanced T1-weighted imaging and DWI at 1.5 T. Read-out segmented multi-shot DWI was acquired at b values of 0 s/mm 2 , 200 s/mm 2 , 400 s/mm 2 and 800 s/mm 2 . We calculated the IVIM parameters perfusion fraction (f) and tissue diffusion coefficient (D). Diffusion-weighted images at b=800 s/mm 2 , f parameter maps and post-contrast T1-weighted images were retrospectively assessed by two independent readers for synovitis using the Juvenile Arthritis MRI Scoring system., Results: Seven (58%) children showed synovial hypertrophy on contrast-enhanced imaging. Diagnostic ratings for synovitis on DWI and on f maps were fully consistent with contrast-enhanced imaging, the diagnostic reference. Two children had equivocal low-confidence assessments on DWI. Median f was 6.7±2.0% for synovitis, 2.1±1.2% for effusion, 5.0±1.0% for muscle and 10.6±5.7% for popliteal lymph nodes. Diagnostic confidence was higher based on f maps in three (25%) children and lower in one child (8%), as compared to DWI., Conclusion: DWI with IVIM reliably visualises synovitis of the knee joint. Perfusion fraction maps differentiate thickened synovium from joint effusion and hence increase diagnostic confidence.

By combining intravoxel incoherent motion (IVIM) and diffusion tensor imaging (DTI) we introduce a new diffusion model called intravoxel oriented flow (IVOF) that accounts for anisotropy of diffusion and the flow-related signal. An IVOF model using a simplified apparent flow fraction tensor (IVOFf ) is applied to diffusion-weighted imaging of human kidneys. The kidneys of 13 healthy volunteers were examined on a 3 T scanner. Diffusion-weighted images were acquired with six b values between 0 and 800 s/mm(2) and 30 diffusion directions. Diffusivity and flow fraction were calculated for different diffusion models. The Akaike information criterion was used to compare the model fit of the proposed IVOFf model to IVIM and DTI. In the majority of voxels the proposed IVOFf model with a simplified apparent flow fraction tensor performs better than IVIM and DTI. Mean diffusivity is significantly higher in DTI compared with models that account for the flow-related signal. The fractional anisotropy of diffusion is significantly reduced when flow fraction is considered to be anisotropic. Anisotropy of the apparent flow fraction tensor is significantly higher in the renal medulla than in the cortex region. The IVOFf model describes diffusion-weighted data in the human kidney more accurately than IVIM or DTI. The apparent flow fraction in the kidney proved to be anisotropic., (Copyright © 2016 John Wiley & Sons, Ltd.)

Human campylobacteriosis is primarily associated with poultry but also cattle. In this study, 55 Campylobacter jejuni strains isolated from 382 dairy calves' feces were differentiated by multilocus sequence typing and tested for antimicrobial resistance. The most prevalent sequence type (ST) was ST883 (20.0%), followed by ST48 (14.5%), and ST50 (9.1%). In contrast to ST48 and ST50, ST883 has rarely been described in cattle previously. Furthermore, risk factor analysis was performed for the presence of the most prevalent STs in these calves. Multiple regression analysis revealed that the type of farm (organic vs. conventional) and calf housing (place, and individual vs. group) were identified as significantly (p < 0.05) associated with the presence of ST883 in calves, and ST50 was associated with calf diarrhea. Antimicrobial resistance was detected in 58.2% of the isolates. Most of the resistant isolates (81.3%) were resistant to more than one antimicrobial. Most frequently, resistance to ciprofloxacin (49.1%), followed by nalidixic acid (42.8%), and tetracycline (14.5%) was observed. The results of the present study support the hypothesis that dairy calves may serve as a potential reservoir for C. jejuni and pose a risk for transmission, including antimicrobial resistant isolates to the environment and to humans.

Campylobacteriosis is the most important bacterial food-borne disease in the developed world. Consumption of chicken meat, beef or raw milk, direct contact with ruminants and exposure to contaminated surface water or even consumption of tap water have been identified as risk factors for human disease. However, the most important risk factor is consumption of and/or handling contaminated chicken. Campylobacter spp. are fastidious microorganisms but must somehow survive outside the host, especially in food and agricultural environments and also resist the innate and humoral immune responses inside the host. In this paper we hypothesize that other microorganisms in mixed populations with Campylobacter may act to improve survival outside the host and may also protect the pathogen against the intestinal immune system. Our evidence for this hypothesis is based on: 1. newly generated microbial community analysis; 2. the prolonged survival of Campylobacter in mixed species biofilms and in co-culture with environmental bacteria; 3. improved survival in amoebae and rumen fluid; 4. sulfur release and iron uptake systems within the intestinal lumen. This would make Campylobacter an exceptional food-borne pathogen. With this in mind, new strategies are necessary to combat Campylobacter along the total food chain.

Antimicrobial resistance in microbes poses a global and increasing threat to public health. The horizontal transfer of antimicrobial resistance genes was thought to be due largely to conjugative plasmids or transposons, with only a minor part being played by transduction through bacteriophages. However, whole-genome sequencing has recently shown that the latter mechanism could be highly important in the exchange of antimicrobial resistance genes between microorganisms and environments. The transfer of antimicrobial resistance genes by phages could underlie the origin of resistant bacteria found in food. We show that chicken meat carries a number of phages capable of transferring antimicrobial resistance. Of 243 phages randomly isolated from chicken meat, about a quarter (24.7%) were able to transduce resistance to one or more of the five antimicrobials tested into Escherichia coli ATCC 13706 (DSM 12242). Resistance to kanamycin was transduced the most often, followed by that to chloramphenicol, with four phages transducing tetracycline resistance and three transducing ampicillin resistance. Phages able to transduce antimicrobial resistance were isolated from 44% of the samples of chicken meat that we tested. The statistically significant (P = 0.01) relationship between the presence of phages transducing kanamycin resistance and E. coli isolates resistant to this antibiotic suggests that transduction may be an important mechanism for transferring kanamycin resistance to E. coli. It appears that the transduction of resistance to certain antimicrobials, e.g., kanamycin, not only is widely distributed in E. coli isolates found on meat but also could represent a major mechanism for resistance transfer. The result is of high importance for animal and human health., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

In most European countries human campylobacteriosis is the most important bacterial zoonotic foodborne infection. Chicken meat is considered the main source of infection. Since most strategies assessed so far, in reducing Campylobacter colonization in chickens or in the reduction of human disease, have not been very effective, new knowledge regarding Campylobacter's interaction with the host is needed. In this study we analysed fecal and cecal samples of five chicken flocks of different Austrian farms for the occurrence of Cjejuni and C. coli, and analysed the intestinal microbiota by PCR-SSCP, cultural detection of lactic acid bacteria, Enterococci, Staphylococci, Enterobacteriaceae, E. coli, and total aerobic colony counts. Furthermore ten chicken samples of cecal content of a flock during colonization with Campylobacter spp. was analysed by high throughput sequencing. With all three methods used we could detect a change within the microbiota caused by Cjejuni. Enumeration of different bacteria was significantly lower in fecal samples positive for C. jejuni, pointing out that a higher water content and thus, a preliminary stage of diarrhea might appear during Campylobacter colonization. By PCR-SSCP analysis the microbiota composition differed between colonized and non-colonized chicken fecal samples. This could also be detected in community analysis by high throughput sequencing, but this difference was only a tendency and not statistically significant. It can be concluded that C. jejuni is interacting with the intestinal microflora in their respective hosts and hence, this has to be taken into account when providing new strategies to combat Campylobacter colonization and disease.

Membrane peptides appear as an emerging class of regulatory molecules in bacteria, which can interact with membrane proteins, including transporters and sensor kinases. The KdpF peptide, which is cotranscribed with kdpABC genes and regulated by the KdpDE two-component system, is supposed to stabilize the KdpABC potassium transporter complex but may also exhibit unsuspected regulatory function(s). The mycobacterial KdpF can interact with the KdpD histidine kinase, and kdpF overexpression has been shown to reduce intramacrophage replication of Mycobacterium bovis BCG. In this study, we investigated whether KdpF displays similar behavior in another intracellular pathogen, Salmonella enterica serovar Typhimurium. We show that Salmonella KdpF can interact with KdpD in a bacterial two-hybrid assay. We have constructed a Salmonella strain overexpressing kdpF, and we have investigated expression of the kdp regulon, as well as intramacrophage survival. We show that kdpF overexpression reduces expression of kdpA and kdpD genes under potassium limitation. Moreover, kdpF overexpression increases intramacrophage multiplication of S. Typhimurium. Hence, our results indicate that KdpF can play a regulatory role in S. Typhimurium, modulating kdp gene expression and intramacrophage survival, but in a way that differs from the one reported for M. bovis BCG., (© 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.)

Purpose: Phase Contrast Magnetic Resonance Imaging (MRI) is a tool for non-invasive determination of flow velocities inside blood vessels. Because Phase Contrast MRI only measures a single mean velocity per voxel, it is only applicable to vessels significantly larger than the voxel size. In contrast, Fourier Velocity Encoding measures the entire velocity distribution inside a voxel, but requires a much longer acquisition time. For accurate diagnosis of stenosis in vessels on the scale of spatial resolution, it is important to know the velocity distribution of a voxel. Our aim was to determine velocity distributions with accelerated Fourier Velocity Encoding in an acquisition time required for a conventional Phase Contrast image., Materials and Methods: We imaged the femoral artery of healthy volunteers with ECG-triggered, radial CINE acquisition. Data acquisition was accelerated by undersampling, while missing data were reconstructed by Compressed Sensing. Velocity spectra of the vessel were evaluated by high resolution Phase Contrast images and compared to spectra from fully sampled and undersampled Fourier Velocity Encoding. By means of undersampling, it was possible to reduce the scan time for Fourier Velocity Encoding to the duration required for a conventional Phase Contrast image., Results: Acquisition time for a fully sampled data set with 12 different Velocity Encodings was 40 min. By applying a 12.6-fold retrospective undersampling, a data set was generated equal to 3:10 min acquisition time, which is similar to a conventional Phase Contrast measurement. Velocity spectra from fully sampled and undersampled Fourier Velocity Encoded images are in good agreement and show the same maximum velocities as compared to velocity maps from Phase Contrast measurements., Conclusion: Compressed Sensing proved to reliably reconstruct Fourier Velocity Encoded data. Our results indicate that Fourier Velocity Encoding allows an accurate determination of the velocity distribution in vessels in the order of the voxel size. Thus, compared to normal Phase Contrast measurements delivering only mean velocities, no additional scan time is necessary to retrieve meaningful velocity spectra in small vessels., (Copyright © 2013. Published by Elsevier GmbH.)

A Gram-stain-positive, pleomorphic, oxidase-negative, non-motile isolate from the skin of a dog, designated strain 410(T), was subjected to comprehensive taxonomic characterization. Comparison of the 16S rRNA gene sequences revealed that the novel isolate showed highest similarities to the type strains of Corynebacterium humireducens, Corynebacterium diphtheriae, Corynebacterium pseudotuberculosis and Corynebacterium ulcerans (96.1-96.8 %). The quinone system consisted predominantly of MK-8(H(2)) and MK-9(H(2)). The polar lipid profile of strain 410(T) contained the major compounds diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, two unidentified phospholipids and four unidentified glycolipids. The polyamine pattern was composed of the major amines spermidine and spermine. In the fatty acid profile, predominantly straight-chain, saturated and mono-unsaturated fatty acids were detected (C(18 : 1)ω9c, C(16 : 1)ω7c, C(16 : 0)). These chemotaxonomic traits are in agreement with those reported for representatives of the genus Corynebacterium. Strain 410(T) tested negative for diphtheria toxin. Physiological properties as well as unique traits in the polar lipid profile could be used to distinguish strain 410(T) from the most closely related species. These data suggest that strain 410(T) represents a novel species of the genus Corynebacterium, for which we propose the name Corynebacterium epidermidicanis sp. nov. The type strain is 410(T) (= DSM 45586(T) = LMG 26322(T) = CCUG 60915(T)).

Campylobacter jejuni is a major cause of the Guillain-Barré syndrome (GBS) and related diseases. These autoimmune diseases are caused by antibodies cross-reacting with the peripheral (GBS) and central neural tissue (Miller Fisher syndrome - MFS, Bicker-staff's brainstem encephalitis - BBE), leading to acute polyneuropathy. Recently, specific gene loci in C. jejuni have been distinguished which are associated with the onset of GBS, despite a molecular or phenotypic clustering. In this study, we used PCR to analyse C. jejuni isolates of different origin (i.e. bovine, poultry, human) for these genes. A total of 196 isolates were tested for cst-II and neuA. Of these, 101 isolates harboured the cst-II locus and 102 the neuA locus. Eighty-six isolates (44%) hold both genes. The frequency of cst-II in different sources of isolates of bovine, poultry and human isolates did not vary significantly (52, 50 and 52%, respectively). In contrast, the neuA locus was less often found in poultry isolates. Two human strains - from a family outbreak of campylobacteriosis (in 1989 in Austria) in which one person developed MFS - harboured both genes. Thus, although only one in more than 3000 patients with Campylobacter-associated enteritis develop GBS, about half of Campylobacter jejuni strains found in different environments are possibly able to cause GBS. These strains almost equally distributed in bovine, poultry and human isolates. Our results suggest that isolates associated with GBS are not selected by environmental or host-specific factors. Accordingly, this study indicates that host factors such as humoral and cellular immunity are possibly responsible for the development of these autoimmune diseases.

Background: Campylobacter jejuni is one of the most important agents of zoonotic disease. Production as well as companion animals can be the infectious source for Campylobacteriosis in humans. Hence, epidemiological research on animal colonization, survival in food of animal origin, and human Campylobacteriosis is of high priority. As such studies involve worldwide co-operations and should include further typing of isolates in reference centers, using a reliable method for transportation is essential. In the case of C. jejuni, a pathogenic and microaerophilic bacterium, special safety precautions as well as particular transport conditions that guarantee survival of isolates are required., Objective: The purpose of this study was to test various media and temperatures for the transportation of C. jejuni under aerobic conditions and to identify a cheap, effective and easy method that is appropriate for long distance transportation and can be applied by most veterinary/medical laboratories with a basic infrastructure., Materials and Methods: We examined Mueller-Hinton (MH) agar with and w/o 2% horse blood and m-CCDA at room temperature and 2 ± 2 (SD)°C under atmospheric conditions for survival of Campylobacter strains., Results: MH agar with 2% horse blood, suitable transport vials, and an optimum temperature of 2 ± 2°C provided survival of three Campylobacter type strains for at least one month under atmospheric conditions. This was validated by a transport test in which 101 isolates were shipped from Turkey to Austria. All isolates could be recultured and 97% survived more than one month in the transport medium., Conclusion: These findings indicate that the described approach is suitable for inter-laboratory transport of C. jejuni isolates.

Campylobacter jejuni is a major food-borne pathogen. Despite causing enteritis in humans, it is a well-adapted intestinal microorganism in animals, hardly ever generating disease symptoms. Nevertheless, as a true microaerophilic microorganism it is still puzzling how Campylobacter cells can survive on chicken meat, the main source of human infection. In this study, we demonstrate that C. jejuni is able to withstand conditions of atmospheric oxygen tension when cocultured with Pseudomonas species, major food-spoiling bacteria that are frequently found on chicken meat in rather high numbers. Using an in vitro survival assay, interactions of 145 C. jejuni wild-type strains and field isolates from chicken meat, broiler feces, and human clinical samples with type strains and food isolates of Pseudomonas spp., Proteus mirabilis, Citrobacter freundii, Micrococcus luteus, and Enterococcus faecalis were studied. When inoculated alone or in coculture with Proteus mirabilis, Citrobacter freundii, Micrococcus luteus, or Enterococcus faecalis type strains, Campylobacter cells were able to survive ambient oxygen levels for no more than 18 h. In contrast, Campylobacter bacteria inoculated with type strains or wild-type isolates of Pseudomonas showed a prolonged aerobic survival of up to >48 h. This microbial commensalism was diverse in C. jejuni isolates from different sources; isolates from chicken meat and humans in coculture with Pseudomonas putida were able to use this survival support better than fecal isolates from broilers. Scanning electron microscopy revealed the development of fiberlike structures braiding P. putida and C. jejuni cells. Hence, it seems that microaerophilic C. jejuni is able to survive ambient atmospheric oxygen tension by metabolic commensalism with Pseudomonas spp. This bacterium-bacterium interaction might set the basis for survival of C. jejuni on chicken meat and thus be the prerequisite step in the pathway toward human infection.

Controversy exists on veterinary drug application in food animal production and the relevance for human health of antimicrobial resistant commensals isolated from food. The aim of this study was to analyze antimicrobial resistance in Escherichia coli isolated from retail meat of various animal species (including wild roe deer) in Austria. Our results were analyzed taking into consideration the current practices of Austrian veterinarians with regard to their use of antibiotic drugs during pig, poultry, and beef production. Resistant isolates were found most often in pork (76%) followed by poultry (63%) and beef (40%). On wild deer carcasses purchased from Austrian hunters only one isolate was found to be resistant. The latter indicates that antimicrobial resistance is not yet an environmental problem in animals living in the wild. The common use of tetracyclines in veterinary medication in various animal species is clearly reflected in the incidence of antimicrobial resistant isolates in commensal E. coli. The intensive use of fluoroquinolones in poultry could explain the high numbers of nalidixic acid resistant isolates found on poultry meat. Our findings partly explain the impact of veterinary drug application on the resistance development of E. coli isolated from meat.

There is a demand from the meat industry as well as from public health authorities for a simple and rapid detection method for thermophilic Campylobacter spp. from food. Hence, we compared different isolation procedures for their usefulness for this purpose. Bolton enrichment medium without blood, incubated statically in stomacher bags in microaerophilic atmosphere, detected more samples positive for thermophilic Campylobacter spp. than did Preston enrichment broth in bottles with small headspace and tight caps, incubated in aerobic atmosphere. Use of an automated antigen detection system to identify enrichment cultures positive for Campylobacter spp. was as sensitive as selective agars, and reduced the detection time by 24 h. Campylobacter spp. were recovered from 18.4% of the 461 samples tested. The prevalence was highest in refrigerated poultry meat (52% of the 80 samples tested) and poultry offal (41% of the 44 samples tested).

This study was performed to evaluate the resistance rate against antimicrobials of food isolates of the five major food-borne pathogens to compare these and to possibly distinguish a pattern. A total of 922 samples of the major meat species (pork, beef and poultry) were analysed for thermophilic Campylobacter, Salmonella, Yersinia enterocolitica, pathogenic Escherichia coli and Listeria monocytogenes. Isolates were subjected to antimicrobial resistance testing by the disc diffusion method. Roughly the same overall rate of resistance was identified for thermophilic Campylobacter, Salmonella and pathogenic E. coli. Resistance to quinolones and tetracycline was determined most frequently. In contrast, food isolates of Y. enterocolitica and L. monocytogenes were rarely tested resistant. The significance of our findings is that resistance rates in enteric bacteria seem to be much higher than in pathogens found in a variety of environments, closely associated to the host environment.

Yersinia enterocolitica is an important foodborne pathogen, but isolation of virulent Yersinia from food sources is still time consuming and requires skills. In this article, we describe a rapid urease screening on cefsulodin-irgasan-novobiocin (CIN) agar plates with an agar overlay assay. This test is simple to perform, all colonies on a plate can be checked simultaneously, it only takes minutes for detection of urease-positive colonies and the colonies survive for transfer, further characterisation, and storage. Additionally, this method is useful to isolate virulent (urease-positive and pYV harbouring) Y. enterocolitica from foodstuffs.

The Gram-negative pathogen Salmonella enterica harbours a periplasmic D-Ala-D-Ala dipeptidase (termed PcgL), which confers the ability to grow on D-Ala-D-Ala as sole carbon source. We now demonstrate that inactivation of the pcgL gene renders Salmonella hypervirulent. This phenotype results from the accumulation of peptidoglycan-derived D-Ala-D-Ala in the pcgL mutant and not from an intrinsically faster growth rate. Synthetic D-Ala-D-Ala (but not L-Ala-L-Ala or D-Ala) increased the number of wild-type Salmonella in the liver and spleen of mice within 24 h of injection, suggesting that D-Ala-D-Ala interferes with some aspect of innate immunity. However, the pcgL mutant was unable to grow on D-Ala-D-Ala as sole carbon source and was defective for survival in nutrient-poor conditions. We identified clinical isolates lacking D-Ala-D-Ala dipeptidase activity and unable to grow on D-Ala-D-Ala because of inactivation of the pcgL gene. Our data suggest that genes (such as pcgL) that, when mutated make pathogens more virulent, may be retained because their contribution to pathogen fitness in non-host environments outweighs potential advantages of the hypervirulent vari-ant in the infected host.

The VanX protein is a D-alanyl-D-alanine (D-Ala-D-Ala) dipeptidase essential for resistance to the glycopeptide antibiotic vancomycin. While this enzymatic activity has been typically associated with vancomycin- and teicoplainin-resistant enterococci, we now report the identification of a D-Ala-D-Ala dipeptidase in the gram-negative species Salmonella enterica. The Salmonella enzyme is only 36% identical to VanX but exhibits a similar substrate specificity: it hydrolyzes D-Ala-D-Ala, DL-Ala-DL-Phe, and D-Ala-Gly but not the tripeptides D-Ala-D-Ala-D-Ala and DL-Ala-DL-Lys-Gly or the dipeptides L-Ala-L-Ala, N-acetyl-D-Ala-D-Ala, and L-Leu-Pro. The Salmonella dipeptidase gene, designated pcgL, appears to have been acquired by horizontal gene transfer because pcgL-hybridizing sequences were not detected in related bacterial species and the G+C content of the pcgL-containing region (41%) is much lower than the overall G+C content of the Salmonella chromosome (52%). In contrast to wild-type Salmonella, a pcgL mutant was unable to use D-Ala-D-Ala as a sole carbon source. The pcgL gene conferred D-Ala-D-Ala dipeptidase activity upon Escherichia coli K-12 but did not allow growth on D-Ala-D-Ala. The PcgL protein localizes to the periplasmic space of Salmonella, suggesting that this dipeptidase participates in peptidoglycan metabolism.

The influence, on silver staining, of proteins and restriction enzymes in polymerase chain reaction (PCR) products was studied in 12-20% polyacrylamide gels. For small DNA fragments (74, 41 and 33 bp) best results were achieved using 20% polyacrylamide gels. In 12% gels, restriction enzymes and also proteins used for PCR (bovine serum albumin, Taq-DNA polymerase) interfere with silver staining of nucleic acids.